By Alastair Aitken, Michèle Learmonth (auth.), John M. Walker (eds.)
A extraordinary treasury of one hundred forty four cutting-edge and hugely winning analytical innovations for learning proteins and peptides. John Walker has incorporated many up to date and better equipment from his best-selling uncomplicated Protein and Peptide Protocols in addition to including approximately a hundred new ones. each one tried-and-tested protocol includes exact, step by step directions, time-saving troubleshooting suggestions, replacement methods, informative motives, and accomplished lists of reagents and suppliers-aids no longer often present in commonplace magazine recipes. The ideas don't require refined and dear gear.
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Additional info for The Protein Protocols Handbook
95-119. 10. Kirazov, L. , Venkov, L. , and Kirazov, E. P. (1993) Comparison of the Lowry and the Bradford protein assays as applied for protein estimation of membrane-containing fractions. Anal. Biochem. 208, 44--48. 11. Wilson, C. M. (1979) Studies and critique of Amido Black 10B, Coomassie blue R and Fast green FCF as stains for proteins after polyacrylamide gel electrophoresis. Anal. Biochem. 96, 263-278. 12. Sedmak, J. J. and Grossberg, S. E. (1977) A rapid, sensitive and versatile assay for protein using Coomassie brilliant blue G250.
The fluorescence signal from the second population of cells should not be significantly decreased when compared to the first. 7. A swinging bucket microcentrifuge is preferable because fewer cells are lost. 8. Adjust time and temperature for maximum signal-to-noise ratio. Gentle rocking is recommended. 9. The samples should be refiltered if reanalyzed on the flow cytometer as clumps will form. Samples have been reanalyzed up to 1 wk later, but resolution is generally decreased. 10. H. , Westwood, MA) with an air-cooled argon laser (model 532, Omnichrome, Chino, CA).
3. ). 4. Use the hole puncher to excise stained dots from the nitrocellulose membrane, and place them stained side down into blank wells on the microtiter plate (see Note 11). 5. Use the hole puncher to excise blank nitrocellulose circles, and place them in microtiter plate wells containing copper iodide-stained protein and also in some blank wells to determine the background density (see Fig. 1). 6. For transmittance densitometry, first add 5 ~tL/well of household three-in-one oil to make the nitrocellulose translucent, thus reducing the background.